I need help on Ligations

[brett at BORCIM.WUSTL.EDU]

>
>yeah, Jennifer, but I checked the B/M catalog and it said that I did not 
>need to have extra bases. As such, I designed my primer without an extra 
>base. live and Learn right??? So to all, what do I do now.. If truly, the 
>NcoI is not cutting then I am SOL. Any hints...
>

You could: blunt the PCRs with T4 DNAP, Ligate them together into a high
MW mass, then cut with NcoI. A good portion of the new fragments will be cut
where you expected. Anyways, I don't think this is your problem.
See my previous post on this subject.



Brett Lindenbach
    
Lucille P. Markey Student in Human Pathobiology    
Program in Immunology                              
Washington University - St Louis                  
brett at borcim.wustl.edu